Gillespie-Brown J, Fuller S J, Bogoyevitch M A, Cowley S, Sugden P H
Department of Cardiac Medicine, National Heart and Lung Institute, Imperial College of Science, Technology and Medicine, London, United Kingdom.
J Biol Chem. 1995 Nov 24;270(47):28092-6. doi: 10.1074/jbc.270.47.28092.
Adult mammalian ventricular cardiomyocytes are terminally differentiated cells that enlarge adaptively by hypertrophy. In this situation, genes normally expressed in the fetal ventricular cardiomyocyte (e.g. atrial natriuretic factor (ANF), beta-myosin heavy chain (beta-MHC), and skeletal muscle (SkM) alpha-actin) are re-expressed, and there is transient expression of immediate early genes (e.g. c-fos). Using appropriate reporter plasmids, we studied the effects of transfection of the constitutively active or dominant negative mitogen-activated protein kinase kinase MEK1 on ANF, beta-MHC, and SkM alpha-actin promoter activities in cultured ventricular cardiomyocytes. ANF expression was stimulated (maximally 75-fold) by the hypertrophic agonist phenylephrine in a dose-dependent manner (EC50, 10 microM), and this stimulation was inhibited by dominant negative MEK1. Cotransfection of dominant negative MEK1 with a dominant negative mitogen-activated protein kinase (extracellular signal-regulated protein kinase (ERK2)) increased this inhibition. Transfection with constitutively active MEK1 constructs doubled ANF promoter activity. The additional cotransfection of wild-type ERK2 stimulated ANF promoter activity by about 5-fold. Expression of beta-MHC and SkM alpha-actin was also stimulated. Promoter activity regulated by activator protein-1 or c-fos serum response element consensus sequences was also increased. We conclude that the MEK1/ERK2 cascade may play a role in regulating gene expression during hypertrophy.
成年哺乳动物心室心肌细胞是终末分化细胞,可通过肥大进行适应性增大。在这种情况下,通常在胎儿心室心肌细胞中表达的基因(如心钠素(ANF)、β-肌球蛋白重链(β-MHC)和骨骼肌(SkM)α-肌动蛋白)会重新表达,并且即刻早期基因(如c-fos)会有短暂表达。使用合适的报告质粒,我们研究了组成型激活或显性负性促分裂原活化蛋白激酶激酶MEK1转染对培养的心室心肌细胞中ANF、β-MHC和SkMα-肌动蛋白启动子活性的影响。肥大激动剂去氧肾上腺素以剂量依赖性方式(EC50,10 microM)刺激ANF表达(最大75倍),而这种刺激被显性负性MEK1抑制。显性负性MEK1与显性负性促分裂原活化蛋白激酶(细胞外信号调节蛋白激酶(ERK2))共转染可增强这种抑制作用。用组成型激活的MEK1构建体转染使ANF启动子活性增加一倍。野生型ERK2的额外共转染使ANF启动子活性刺激约5倍。β-MHC和SkMα-肌动蛋白的表达也受到刺激。由激活蛋白-1或c-fos血清反应元件共有序列调节的启动子活性也增加。我们得出结论,MEK1/ERK2级联可能在肥大过程中调节基因表达中发挥作用。